Cömez Birnur, Özbaş Suna
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara University, İstanbul 34854, Turkey.
Gels. 2023 Jul 4;9(7):541. doi: 10.3390/gels9070541.
The hydrogels prepared with alginate and chitosan polymers were prepared to deliver the shRNA-encoding plasmid (pshRNA) to MDA-MB-231 cells for the inhibition of β-catenin (CTNNB1), which was reported to be overexpressed in breast cancer. Polyion complex hydrogels prepared using sodium alginate and chitosan were characterized by Fourier transform infrared spectrometry (FTIR) analysis, scanning electron microscope (SEM) analysis, swelling, and degradation properties. After the release properties and serum stability of pshRNA-loaded hydrogels were determined, their cytotoxicity, transfection efficacy, and effects on CTNNB1 expression were investigated in MDA-MB-231 cells. All hydrogels were shown to protect pshRNA from the enzymatic activity of serum and to deliver pshRNA to cells efficiently. As a result of transfection studies, pshRNA-loaded hydrogels reduced CTNNB1 expression by up to 30.25%. Cell viability also decreased by 38% in cells treated with 2.5% (/) alginate-chitosan hydrogel containing pshRNA targeting CTNNB1. Alginate-chitosan hydrogels were shown to be a suitable matrix system for local gene delivery.
制备了由海藻酸盐和壳聚糖聚合物组成的水凝胶，用于将编码短发夹RNA的质粒（pshRNA）递送至MDA-MB-231细胞，以抑制β-连环蛋白（CTNNB1），据报道该蛋白在乳腺癌中过表达。使用海藻酸钠和壳聚糖制备的聚离子复合水凝胶通过傅里叶变换红外光谱（FTIR）分析、扫描电子显微镜（SEM）分析、溶胀和降解特性进行表征。在测定了负载pshRNA的水凝胶的释放特性和血清稳定性后，研究了它们在MDA-MB-231细胞中的细胞毒性、转染效率以及对CTNNB1表达的影响。结果表明，所有水凝胶均能保护pshRNA免受血清酶活性的影响，并能有效地将pshRNA递送至细胞。转染研究结果显示，负载pshRNA的水凝胶可将CTNNB1表达降低多达30.25%。在用靶向CTNNB1的2.5%（/）海藻酸盐-壳聚糖水凝胶处理的细胞中，细胞活力也下降了38%。海藻酸盐-壳聚糖水凝胶被证明是一种适合局部基因递送的基质系统。
